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CO1 Barcode Assay

The cytochrome c oxidase subunit 1 (CO1) gene is an ideal target for species discrimination as this gene is conserved across higher eukaryotes, including insects, fish, birds and mammals. CO1 is located on mitochondrial DNA, meaning it has a relatively fast mutation rate that aligns with speciation timescales.

An approximately 650bp region of CO1 known as the barcode region is amplified with degenerate primers followed by DNA sequence analysis by the Sanger method. For insect species a smaller region of approximately 450bp is targeted on the CO1 gene.

The obtained DNA sequence is compared against a known standard for species confirmation.